Acute phase response in mice experimentally infected with Trypanosoma congolense: a molecular gauge of parasite-host interaction

Mice infected with Trypanosoma congolense developed a severe anaemia 1 week after infection, which persisted till treatment with diminazine aceturate when the packed cell volume (PCV) recovered to pre-infection levels. This was accompanied by a marked increase in the plasma levels of the acute phase proteins (APP), serum amyloid P-component (SAP) and haptoglobin (Hp). The initial peak levels of Hp and SAP were attained 7 and 12 days post-infection (DPI), respectively. Thereafter SAP levels decreased significantly to near pre-infection levels, but later increased even after treatment to give a second peak 34 DPI after which there was a decline till the study was terminated. The Hp levels on the other hand decreased to an intermediate level after the initial peak increasing to a second peak 22 DPI. Thereafter Hp decreased significantly following diminazine aceturate treatment to reach pre-infection levels within 5 days post-treatment. This indicates that T. congolense-infected mice develop severe anaemia accompanied by an acute phase response leading to an increase in SAP and Hp but that following treatment divergent responses occurred indicating differences in the pathways for stimulation of the APP. Haptoglobin was shown to be an earlier indicator of infection and a better marker in monitoring the response to treatment.     

Renal amyloidosis in cattle with inflammatory diseases

BACKGROUND: The association of inflammatory diseases such as traumatic reticuloperitonitis (TRP), mastitis, metritis, and pododermatitis with renal amyloidosis in cattle is poorly described. HYPOTHESIS: Serum amyloid A (SAA) levels are elevated during inflammatory diseases, and renal amyloidosis is formed as a complication. ANIMALS: This study was conducted with 82 crossbred cattle with mastitis (n = 18 cows), metritis (n = 11 cows), TRP (n = 30 cows), and pododermatitis (n = 23 : 15 cows and 8 beef cattle). Ten clinically healthy cows served as controls. Methods: Hematological, urinary, and blood parameters, including SAA, were measured by an automated procedure provided with trade kits. Determination of amyloidal structures was made by histopathological examination of renal biopsy specimens. RESULTS: At the end of this trial, amyloidosis was detected in 5 cows displaying typical nephrotic syndrome, with hypoproteinemia and proteinuria in combination with polyuria and weight loss. Furthermore, it was observed that cows with renal amyloidosis had significantly higher (P < .01) total leukocyte counts, serum and urine enzyme activities, and urea and creatinine concentrations, with lower serum total protein concentrations, when compared with animals without renal amyloidosis. CONCLUSIONS AND CLINICAL IMPORTANCE: The incidence of AA amyloidosis in cattle in this study suggests that cattle with mastitis, metritis, and pododermatitis have a high prevalence of systemic amyloidosis in response to inflammation.     

昆虫蛋白质饲料在动物生产中的应用

昆虫是最具开发潜力的动物性蛋白质饲料资源。作者就昆虫的营养价值、昆虫蛋白质饲料在畜牧业生产上的开发应用效果进行了综述,最后提出了昆虫饲料的发展前景。     

鸭肠炎病毒NP蛋白的原核表达及间接ELISA方法的建立

设计一对特异性引物扩增出鸭肠炎病毒(DEV)核衣壳蛋白(NP)基因,并将其定向插入到原核表达载体pET32a上,构建了NP基因的原核表达载体pET-NP;将重组载体pET-NP转化表达宿主菌BL21后,经SDS-PAGE分离后行Western blot显示,获得的表达产物具有良好的免疫原性;应用His.Bind亲和层析柱纯化重组NP蛋白,并以此作为包被抗原,初步建立了检测鸭肠炎病毒抗体的iNP-ELISA;经方阵滴定确定,重组蛋白抗原的最佳包被浓度为5.0μg/L,血清最佳稀释度为1∶80,阳性判定标准为:待检血清OD405值≥1.2,且待检血清OD405和阴性血清OD405的比值≥2.0;应用iNP-ELISA对450份鸭血清样本进行检测,结果iNP-ELISA与全病毒包被的iDEV-ELISA符合率达90.9%。     

牛杀菌／通透性增加蛋白氮端基因的克隆和序列分析

本试验应用RT-PCR技术,参照Genbank报道的序列,从荷斯坦牛中性粒细胞mRNA中扩增出杀菌／通透性增加蛋白（BPI）氮端基因（713bp）,并与pGEM-T-easy载体连接,构建基因重组体pGEM-T-easy-BPI,进行序列测定。结果表明获得长度为713bp的BPI氮端基因。序列分析证实该片段与安哥斯牛BPI氮端基因相比有1个点突变,为同义突变。该基因的克隆为进一步表达该基因奠定了基础。     

Serum biomarker profiling in cancer studies: a question of standardisation?

Companion animals are exposed to similar environmental conditions and carcinogens as humans. In some animal cancers, there also appears to be the same genetic changes associated as in humans. However, little work has been carried out in cancer biomarker identification in animals. The recent dramatic advances in molecular medicine, genomics, proteomics and translational research will allow biomarker identification, which may provide the best strategies for veterinarians and clinicians to combat disease by early diagnosis and administration of effective treatments. Proteomics may have important applications in cancer diagnosis, prognosis and predictive clinical outcome that could directly change clinical practice by affecting critical elemen‐ts of care and management. This review summarizes the advances in proteomics that has propelled us to this exciting age of clinical proteomics, and highlights the future work that is required for this to become a reality. In this review, we will discuss the available proteomic technologies and their limitations, and highlight the key areas of research and how they have been used to discover cancer biomarkers. The principles described here are equally applicable to human and animal disease, but implementation of ‘omic’ technologies requires stringent guidelines for collection of clinical material, the application of analytical techniques and interpretation of the data.     

柔嫩艾美耳球虫棒状体颈部蛋白2基因真核表达载体的构建及在293T细胞中表达

为构建柔嫩艾美耳球虫棒状体颈部蛋白2(Eimeria tenella rhoptry neck protein 2,EtRON2)基因的重组质粒pCAGGS-EtRON2,并转染293T细胞进行表达,以柔嫩艾美耳球虫孢子化卵囊cDNA为模板,经PCR扩增其核心编码区的一部分,将其克隆至pGEM-Teasy载体,构建pGEM-Teasy-EtRON2质粒,双酶切出目的片段后与相应酶切的真核表达载体pCAGGS连接,构建真核表达质粒pCAGGS-EtRON2。该重组质粒经酶切和测序鉴定后转染293T细胞进行表达,分别用免疫印迹和间接免疫荧光鉴定EtRON2基因的表达情况。所构建的真核表达质粒pCAGGS-EtRON2经过双酶切鉴定,可见一条大小约为1172 bp的目的条带,测序结果与GenBank所登录序列完全一致;免疫印迹实验可见大小约为43 kDa的目的蛋白条带,间接免疫荧光实验可以检测到特异性红色荧光。研究结果表明已成功构建了EtRON2的真核表达质粒pCAGGS-EtRON2,并在真核细胞中获得表达,为深入研究EtRON2的生物学功能奠定了基础。     

饲粮蛋白质水平对冬毛期水貂胃肠道消化酶活性以及空肠形态结构的影响

本文旨在研究饲粮蛋白质水平对冬毛期水貂胃肠道消化酶活性以及空肠形态结构的影响。选择 ９０日龄的健康公貂 ６０只,随机分成 ６组（每组 １０个重复,每个重复 １只）,分别饲喂蛋白质水平为 ２８％（Ⅰ组）、３０％（Ⅱ组）、３２％（Ⅲ组）、３４％（Ⅳ组）、３６％（Ⅴ组）和 ３８％（Ⅵ组）的试验饲粮。预试期 ７ｄ,正试期 １１５ｄ。饲养试验结束后进行屠宰,制备胃肠道内容物上清液,用于测定消化酶活性;制备空肠组织切片,用于观察空肠的形态结构。结果表明：Ⅴ组水貂的胃蛋白酶活性极显著高于Ⅰ、Ⅱ和Ⅲ组（Ｐ＜０．０１）,与Ⅳ和Ⅵ组差异不显著（Ｐ＞０．０５）。Ⅳ组水貂的十二指肠和空肠胰淀粉酶活性极显著高于Ⅰ、Ⅱ、Ⅲ、Ⅴ和Ⅵ组（Ｐ＜０．０１）。Ⅳ和Ⅴ组水貂的回肠胰淀粉酶活性显著或极显著高于Ⅰ和Ⅱ组（Ｐ＜０．０５或 Ｐ＜０．０１）。Ⅳ组水貂的十二指肠胰蛋白酶活性极显著高于Ⅰ、Ⅱ、Ⅴ和Ⅵ组（Ｐ＜０．０１）,空肠胰蛋白酶活性极显著高于Ⅰ、Ⅱ、Ⅲ、Ⅴ和Ⅵ组（Ｐ＜０．０１）。Ⅲ组水貂的回肠胰蛋白酶活性显著高于Ⅵ组（Ｐ＜０．０５）,与其他各组差异不显著（Ｐ＞０．０５）。Ⅵ组水貂的十二指肠胰脂肪酶活性显著或极显著高于Ⅱ、Ⅳ和Ⅴ组（Ｐ＜０．０５或 Ｐ＜０．０１）,空肠胰脂肪酶活性显著或极显著高于Ⅰ、Ⅱ、Ⅲ、Ⅳ和Ⅴ组（Ｐ＜０．０５或 Ｐ＜０．０１）。Ⅰ、Ⅲ和Ⅴ组水貂的回肠胰脂肪酶活性极显著高于Ⅱ和Ⅵ组（Ｐ＜０．０１）,与Ⅳ组差异不显著（Ｐ＞０．０５）。饲粮蛋白质水平为 ３６％时,水貂空肠绒毛高度、隐窝深度、绒毛高度与隐窝深度比值、黏膜厚度以及肠壁厚度均处在较高水平。由此得出,饲粮蛋白质水平可影响冬毛期水貂胃肠道消化酶活性,并可调节其空肠形态结构。     

希腊式酸奶的研制

研制希腊式酸奶,确定希腊式酸奶的最佳配方为鲜牛奶40%、白砂糖8%、脱脂奶粉：WPC为80：20、酪蛋白酸钠2‰、菌种为YO-MIXTM495;同时确定了制作希腊式酸奶的最佳水合条件为50℃、60min,发酵终点pH值控制在4.65。